Mouse IL-6 ELISA Kit - Quantikine
R&D Systems, part of Bio-Techne | Catalog # M6000B
2 plate pack size in process of discontinuation
Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (50 µL), Heparin Plasma (50 µL)
Sensitivity
1.8 pg/mL
Assay Range
7.8-500 pg/mL (Cell Culture Supernates, Serum, Heparin Plasma)
Product Summary for Mouse IL-6 Quantikine ELISA Kit
The Quantikine Mouse IL-6 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse IL-6 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mouse IL-6 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant mouse IL-6. Results obtained using natural mouse IL-6 showed dose response curves that were parallel to the standard curves obtained using the Quantikine mouse kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse IL-6.
Product Specifications
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Reactivity
Mouse
Specificity
Natural and recombinant mouse IL-6
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Sample Values
Serum - Twenty individual mouse serum samples were evaluated for the presence of mouse
IL-6 in this assay. In all samples, IL-6 levels measured below the low standard, 7.8 pg/mL.
A mouse was injected with 15 μg of LPS and was bled 2 hours after injection. The serum was
tested for mouse IL-6 and measured 31,800 pg/mL.
Heparin Plasma - Eight individual mouse heparin plasma samples were evaluated for the
presence of mouse IL-6 in this assay. Six samples were below the low standard, 7.8 pg/mL.
Two samples measured 23 pg/mL and 18 pg/mL.
Cell Culture Supernates:
EL-4 mouse lymphoblast cells (1 x 106
cells/mL) were cultured for 3 days in DMEM
supplemented with 10% fetal bovine serum and stimulated with 10 μg/mL PHA and 10 μg/mL
PMA. The culture supernate was assayed for mouse IL-6 and measured 386 pg/mL.
D10.G4.1 mouse helper T cells (1 x 105
cells/mL) were cultured in RPMI 1640 supplemented
with L-glutamine, 10% fetal bovine serum, 50 μM beta-mercaptoethanol, and 2.5% rat T cell
polyclone media (Collaborative # 40115). During the final 3 days of culture, cells were
stimulated with 2.5 ng/mL LPS. The culture supernate was assayed for mouse IL-6 and
measured 759 pg/mL.
Mouse heart cells were cultured for 7 days in RPMI supplemented with 10% fetal bovine serum.
The culture supernate was assayed for mouse IL-6 and measured 1120 pg/mL.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, Heparin Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 28 | 22 | 27 |
| Mean (pg/mL) | 30 | 87 | 232 | 29 | 85 | 238 |
| Standard Deviation | 2.0 | 3.4 | 8.1 | 1.8 | 7.5 | 18.1 |
| CV% | 6.7 | 3.9 | 3.5 | 6.1 | 8.9 | 7.6 |
Recovery for Mouse IL-6 Quantikine ELISA Kit
The recovery of mouse IL-6 spiked to three levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Supernates (n=5) | 110 | 86-120 |
| Heparin Plasma (n=6) | 101 | 81-115 |
| Serum (n=5) | 99 | 84-113 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of mouse IL-6 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Mouse IL-6 Quantikine ELISA Kit
Mouse IL-6 ELISA Standard Curve
Mouse IL-6 ELISA Kit - Quantikine
Increased production of proinflammatory cytokines and VEGF in the plasma and chondrocytes in high fat diet (HFD)-feeding mice.(a~d) levels of plasma IL-6 (a), TNF-alpha (b), IL-1 beta (c) and VEGF (d) in the C57BL/6 male mice fed with a normal diet (ND) or HFD for 8 weeks. (n = 5, **P < 0.01). (e~h) Relative mRNA expression of IL-6 (e), TNF-alpha (f), IL-1 beta (g) and VEGF (h) in the chondrocytes isolated from the 8-week ND or HFD-feeding mice. (n = 5, **P < 0.01). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26271607), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Mouse IL-6 Quantikine ELISA Kit by ELISA
PYC suppressed LPS-induced proinflammatory cytokine production in BV2 microglia.Cells were incubated with the indicated concentrations of PYC or vehicle for 1h before LPS treatment (500ng/mL). After 24 h incubation, the culture supernatants were collected, and the amount of TNF-alpha, IL-6, IL-1 beta and IL-10 were measured by ELISA (A-D). Data are represented as mean ± SD from at least 3 independent experiments. *P< 0.05 compared with LPS alone. #P< 0.05. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26367267), licensed under a CC-BY license. Not internally tested by R&D Systems.Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: IL-6
IL-6 induces signaling through a cell surface heterodimeric receptor complex composed of a ligand binding subunit (IL-6 R alpha) and a signal transducing subunit (gp130). IL-6 binds to IL-6 Ra, triggering IL-6 Ra association with gp130 and gp130 dimerization. gp130 is also a component of the receptors for CLC, CNTF, CT-1, IL-11, IL-27, LIF, and OSM. Soluble forms of IL-6 Ra are generated by both alternative splicing and proteolytic cleavage. In a mechanism known as trans-signaling, complexes of soluble IL-6 and IL-6 Ra elicit responses from gp130-expressing cells that lack cell surface IL-6 Ra. Trans-signaling enables a wider range of cell types to respond to IL-6, as the expression of gp130 is ubiquitous, while that of IL-6 Ra is predominantly restricted to hepatocytes, monocytes, and resting lymphocytes. Soluble splice forms of gp130 block trans-signaling from IL-6/IL-6 Ra but not from other cytokines that use gp130 as a co-receptor.
IL-6, along with TNF-a and IL-1, drives the acute inflammatory response. IL-6 is almost solely responsible for fever and the acute phase response in the liver, and it is important in the transition from acute inflammation to either acquired immunity or chronic inflammatory disease. When dysregulated, it contributes to chronic inflammation in conditions such as obesity, insulin resistance, inflammatory bowel disease, arthritis, and sepsis. IL-6 modulates bone resorption and is a major effector of inflammatory joint destruction in rheumatoid arthritis through its promotion of Th17 cell development and activity. It contributes to atherosclerotic plaque development and destabilization as well as the development of inflammation-associated carcinogenesis.
Long Name
Interleukin 6
Alternate Names
BSF-2, BSF2, IFNB2, IL6, MGI-2A
Entrez Gene IDs
Gene Symbol
IL6
Additional IL-6 Products
Product Documents for Mouse IL-6 Quantikine ELISA Kit
Product Specific Notices for Mouse IL-6 Quantikine ELISA Kit
For research use only
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⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.